Navigation Links
Mycobacterium smegmatis

Multiporator / Electroporator 2510 Transformation Protocol Protocol No. 4308 915.522 03/2002 Microorganism Mycobacterium smegmatis LR222 Cell type Bacteria, gram positive Molecules injected Plasmid DNA Growth medium Middlebrook 7H9 medium with 0.1% Tween and Dubos oleic albumin complex
enrichment Washing solution 10% glycerol Electroporation solution 10% glycerol Outgrowth medium Middlebrook 7H9 liquid medium with 0.1% Tween and Dubos oleic albumin complex
enrichment, selective Middlebrook 7H10 agar plates Cuvette 1 mm gap width Reference Beggs, M. L.. et al 1995 Journal of Bacteriology 177, No. 17 4836-4840 Making electrocompetent cells:

1. Grow cells in sidearm flask or tissue culture plates with gentle agitation until a Klett unit reading of 100 to 200 was
obtained. 2. Harvest cells and wash three times with cold sterile 10% glycerol. 3. Resuspend 1/100 volume of 10% glycerol.

Electroporation of cells:

  1. Add up to 1 g plasmid DNA to 60 l of electrocompetent cells. Homogenize by gently mixing with pipette several
    times. Incubate on ice for 10 min. Transfer mixture into a prechilled cuvette.
  2. Wipe moisture from the cuvette and insert the cuvette into the device.
  3. Electroporation:

    Mode Prokaryotes O Voltage (V) 1,200 V Time constant (T) 5 ms
  4. Wash cells out of the cuvette with 1 ml of outgrowth medium and incubate for 2 h at 37 C.
  5. Plate on selective agar.
Expected Results: Transformation efficiency up to 105 transformants/g of DNA.


'"/>

Source:


Page: All 1 2

Related biology technology :

1. Mycobacterium avium
2. Mycobacterium intracellulare
3. Mycobacterium smegmatis
4. Mycobacterium tuberculosis
5. Mycobacterium avium
6. Mycobacterium intracellulare
7. Mycobacterium tuberculosis
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:6/24/2016)... on a range of subjects including policies, debt and investment ... Speaking at a lecture to the Canadian Economics ... the country,s inflation target, which is set by both the ... "In certain areas there needs to be frequent ... not sit down and address strategy together?" He ...
(Date:6/23/2016)... WA (PRWEB) , ... June 23, 2016 , ... ... announces the release of its second eBook, “Clinical Trials Patient Recruitment and Retention ... recruitment and retention in this eBook by providing practical tips, tools, and strategies ...
(Date:6/23/2016)... SPRING, Md. , June 23, 2016 A ... collected from the crime scene to track the criminal down. ... and the U.S. Food and Drug Administration (FDA) uses DNA ... Sound far-fetched? It,s not. The ... genome sequencing to support investigations of foodborne illnesses. Put as ...
(Date:6/23/2016)... , June, 23, 2016  The Biodesign Challenge ... envision new ways to harness living systems and biotechnology, ... Art (MoMA) in New York City ... 130 participating students, showcased projects at MoMA,s Celeste Bartos ... Paola Antonelli , MoMA,s senior curator of architecture and ...
Breaking Biology Technology:
(Date:4/28/2016)... , April 28, 2016 First quarter ... (139.9), up 966% compared with the first quarter of 2015 ... totaled SEK 589.1 M (loss: 18.8) and the operating margin was ... (loss: 0.32) Cash flow from operations was SEK 249.9 ... 2016 revenue guidance is unchanged, SEK 7,000-8,500 M. The ...
(Date:4/19/2016)... DUBAI , UAE, April 20, 2016 ... can be implemented as a compact web-based "all-in-one" system ... in the biometric fingerprint reader or the door interface ... requirements of modern access control systems. The minimal dimensions ... the ID readers into the building installations offer considerable ...
(Date:4/14/2016)... April 14, 2016 BioCatch ... Detection, today announced the appointment of Eyal Goldwerger ... role. Goldwerger,s leadership appointment comes at a ... of the deployment of its platform at several of ... technology, which discerns unique cognitive and physiological factors, is ...
Breaking Biology News(10 mins):